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  Method of separating unattached raman-active tag from bioassay or other reaction mixtureUSPTO Application #: 20060134806Title: Method of separating unattached raman-active tag from bioassay or other reaction mixture Abstract: A method of separating a Raman-active tag unattached to a target from a Raman-active complex is disclosed. The Raman-active complex includes a Raman-active tag attached to a target. The method includes providing a mixture including at least one Raman-active tag unattached to a target and at least one Raman-active complex; and passing the mixture through a porous membrane. (end of abstract) Agent: General Electric Company Global Research - Niskayuna, NY, US Inventor: Frank John Mondello USPTO Applicaton #: 20060134806 - Class: 436524000 (USPTO) Related Patent Categories: Chemistry: Analytical And Immunological Testing, Involving An Insoluble Carrier For Immobilizing Immunochemicals, Carrier Is Inorganic The Patent Description & Claims data below is from USPTO Patent Application 20060134806. Brief Patent Description - Full Patent Description - Patent Application Claims
BACKGROUND OF THE INVENTION [0001] The invention relates to Raman-active bioassays or other reaction mixture. Particularly, the invention is directed to a method of separating an unattached Raman-active tag from a bioassay or other reaction mixture. DESCRIPTION OF RELATED ART [0002] Raman-active tags 100 are known to detect the presence of pathogenic organisms or other materials. FIG. 1 is a schematic representation of a Raman-active tag 100 that includes a Raman-active particle 110 and one or more target-binding moieties 112. The target-binding moiety 112 on the Raman-active tag 100 is configured to allow the Raman-active tag 100 to attach to one or more targets 212 to form a Raman-active complex 200. In contrast, the Raman-active tag 100 is unattached to a target 212. FIG. 2 and FIG. 3 are schematic representations of a Raman-active complex 200 comprising a Raman-active tag 100 and a target 212. In the presence of a target 212, one or more target-binding moieties 112 against a target 212 allow the Raman-active tag 100 to attach to the target 212. Detection of the target 212 is then based on the presence of a Raman signal after removing any Raman-active tags 100 that are unattached to a target 212 from the test mixture. Failure to eliminate unattached Raman-active tags 100 results in false positive detection of the presence of the target 212. Centrifugation is a method commonly used to separate unattached Raman-active tags 100 from Raman active complexes 200 that are attached to a target; however, centrifugation is inefficient because the Raman-active tags 100 have a density such that the Raman-active tags 100 pellet along with the Raman active complexes 200 and targets 212. [0003] Thus, a need still remains for a method of separating unattached Raman-active tag 100 from bioassay or other reaction mixture. SUMMARY [0004] The purpose and advantages of embodiments of the invention will be set forth and apparent from the description that follows, as well as will be learned by practice of the embodiments of the invention. Additional advantages will be realized and attained by the methods and systems particularly pointed out in the written description and claims hereof, as well as from the appended drawings. An embodiment of the invention provides a method of separating a Raman-active tag unattached to a target from a Raman-active complex. The Raman-active complex comprises a Raman-active tag attached to a target. The method comprises providing a mixture comprising at least one Raman-active tag unattached to a target and at least one Raman-active complex; and passing the mixture through a porous membrane. Each Raman-active complex has a first size and the membrane has pores with an effective size smaller than the first size. [0005] A second embodiment provides a method of separating a Raman-active tag unattached to a target from a Raman-active complex. The Raman-active complex comprises a Raman-active tag attached to a target. The method comprises providing a mixture comprising at least one Raman-active tag unattached to a target and at least one Raman-active complex; passing the mixture through a porous membrane; and applying a sufficient force to the porous membrane to separate at least one Raman-active complex from the membrane while the Raman-active tag unattached to a target remains on the membrane. Each Raman-active complex has a first size and the membrane has pores with an effective size smaller than the first size. [0006] A third embodiment provides a method of separating a Raman-active tag unattached to a target from a Raman-active complex. The Raman-active complex comprises a Raman-active tag attached to a target. The method comprises providing a mixture comprising at least one Raman-active tag unattached to a target and at least one Raman-active complex; and passing the mixture through a porous membrane comprising pores, wherein the porous membrane is configured to collect at least one Raman-active complex. Each Raman-active complex has a first size. [0007] The accompanying figures, which are incorporated in and constitute part of this specification, are included to illustrate and provide a further understanding of the method and system of the invention. Together with the description, the drawings serve to explain the principles of the invention. BRIEF DESCRIPTION OF THE DRAWINGS [0008] FIG. 1 is a schematic representation of an unattached Raman-active tag in accordance with an embodiment of the invention; [0009] FIG. 2 is a schematic representation of a Raman-active complex in accordance with an embodiment of the invention; [0010] FIG. 3 is a schematic representation of Raman-active complex in accordance with an embodiment of the invention; [0011] FIG. 4 is a schematic representation of a method of separating a Raman-active tag unattached to a target from a Raman-active complex in accordance with an embodiment of the invention; [0012] FIG. 5 is a flow chart of a method of separating a Raman-active tag unattached to a target from a Raman-active complex in accordance with an embodiment of the invention; [0013] FIG. 6A is a Raman spectrum of a sample of Bacillus spores not exposed to Raman-active tags in accordance with an embodiment of the invention; [0014] FIG. 6B is a Raman spectrum of a sample of Raman-active tags functionalized with anti-bacillus antibodies filtered in the absence of bacillus spores in accordance with an embodiment of the invention; [0015] FIG. 6C is a Raman spectrum of a sample of Bacillus spores exposed to Raman-active tags in accordance with an embodiment of the invention; [0016] FIG. 6D is a Raman spectrum of a sample of Bacillus spores exposed to Raman-active tags designed to detect E. coli cells in accordance with an embodiment of the invention; [0017] FIG. 6E is a Raman spectrum of control samples of the antibody functionalized Raman tags placed directly on a glass surface in accordance with an embodiment of the invention; [0018] FIG. 7A is a Raman spectrum of a sample of E. coli cells not exposed to Raman-active tags in accordance with an embodiment of the invention [0019] FIG. 7B is a Raman spectrum of a sample of Raman-active tags functionalized with anti-E. coli antibodies filtered in the absence of E. coli cells in accordance with an embodiment of the invention; [0020] FIG. 7C is a Raman spectrum of a sample of E. coli cells exposed to Raman-active tags bound to anti-E. coli antibody in accordance with an embodiment of the invention; Continue reading... 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